p21WAF1 induces permanent growth arrest and enhances differentiation, but does not alter apoptosis in PC12 cells
p21WAF1 induces permanent growth arrest and enhances differentiation, but does not alter apoptosis in PC12 cells
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DOI：
10.1038/sj.onc.1201577
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发表时间：
1998-01-29
期刊：
影响因子：
8
通讯作者：
Pittman, RN
中科院分区：
文献类型：
--
作者：
Erhardt, JA;Pittman, RN
p21(WAF1) 细胞周期蛋白依赖性激酶抑制剂与多种细胞系的增殖、分化和死亡的控制有关。为了进一步检查 p21 对这些细胞过程之间转换的调节，将诱导型 p21 载体（lac 操纵子系统）转染到大鼠嗜铬细胞瘤 (PC12) 神经细胞系中，p21 的诱导导致永久生长停滞，细胞计数、FACS 分析和胸苷掺入，即使在去除诱导信号（IPTG）后，这种抑制仍得以维持。 Northern分析显示，IPTG去除后内源性p21 mRNA增加，这可能是导致持续生长停滞的原因，尽管异位p21表达减少，p21过度表达并没有直接导致分化表型；然而，对神经生长因子（NGF）的反应大大加速了分化。为了检查对细胞死亡的影响，并具体测试因不适当进入细胞周期而导致营养支持取消而导致细胞凋亡的假设，从增殖和 p21 停滞的 PC12 细胞中除去血清。细胞凋亡率不受 p21 影响，也不能有效改变其他细胞凋亡刺激后的死亡程度。
p21(WAF1) cyclin-dependent kinase inhibitor has been implicated in the control of proliferation, differentiation, and death in, various cell lines, To further examine p21 regulation of the transitions between these cellular processes, an inducible p21 vector (lac operon system) was transfected into the rat pheochromocytoma (PC12) neural cell line, Induction of p21 led to permanent growth arrest, as evidenced by cell counts, FACS analysis, and thymidine incorporation, This arrest was maintained, even after removal of the inducing signal (IPTG). Northern analysis revealed that endogenous p21 mRNA increased following IPTG removal, which may be responsible for the continued growth arrest despite the decrease in ectopic p21 expression, p21 overexpression did not directly lead to a differentiated phenotype; however, differentiation in response to nerve growth factor (NGF) was greatly accelerated. To examine effects on cell, death, and specifically test the hypothesis that apoptosis caused by withdrawal of trophic support results from inappropriate entry into cell cycle, serum was removed from proliferating and p21-arrested PC12 cells, The rate of apoptotic death was not affected by p21, nor was it effective in altering the extent of death following other apoptotic stimuli.